ObjectiveTo explore the safety and short-term efficacy of uniportal and three-port video-assisted thoracoscopic surgery (VATS) anatomical segmentectomy for pulmonary nodules. MethodsThe clinical data of 225 patients with consecutive VATS anatomic segmentectomy by the same surgeon in Xuzhou Central Hospital between December 2019 and February 2022 was retrospectively reviewed. There were 85 males and 140 females with an average age of 57.3±11.6 years. These patients were divided into an uniportal VATS group (128 patients) and a three-port VATS group (97 patients) according to the surgical procedures. Single-direction anatomical procedure was utilized in the uniportal VATS group. The operation time, blood loss during the surgery, number of dissected lymph nodes, duration and volume of chest drainage, incidence of complications, and postoperative hospital stay of the two groups were compared. ResultsThere was no conversion to thoracotomy, addition of surgical ports, or mortality in this cohort, with tumor-negative surgical margins. The postoperative pathological staining confirmed 2 (0.9%) patients of lymph node metastasis (pN1) and 4 (1.8%) patients of adenocarcinoma with micropapillary component. As compared with the three-port VATS group, patients in the uniportal VATS group had shorter operation time (115.6±54.7 min vs. 141.5±62.8 min, P=0.001), less intraoperative blood loss (77.2±49.6 mL vs. 96.9±98.1 mL, P=0.050), less total thoracic drainage [394.0 (258.8, 580.0) mL vs. 530.0 (335.0, 817.5) mL, P=0.010], and shorter postoperative hospital stay (7.7±3.7 d vs. 8.7±3.5 d, P=0.031). Both groups showed similar stations and numbers of dissected lymph nodes, incidence of operation-related complications, duration of chest tube drainage, and the drainage volume in the first and second postoperative days (P>0.05). No tumor recurrence or metastasis was recorded in this cohort during the follow-up of 11 (1-26) months. ConclusionSingle-direction uniportal VATS anatomical segmentectomy is safe and feasible for the treatment of pulmonary nodules, with better short-term efficacy as compared with the three-port VATS procedure, including shorter operation time, less intraoperative blood loss and thoracic drainage. However, further studies are needed to elucidate the precise indications of segmentectomy for lung cancer.